IGEM:IMPERIAL/2009/Feedback & Debriefs/Feedback 27 7

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Monday 27th July 2009

Module 1

  • Don’t worry about dosage control (will vary on a case by case basis)
  • Do not need tags on the protein - but should add one His tag as proof of concept
  • Enzyme needs to be exactly the same as human enzyme or will this trigger an immune response?

will this be protease resistant?

  • Biobrick protein production for iGEM so we can clone proteins in and out
  • For each application

exactly how to make the protein promoter? E.coli strain? CONTROLS! (measure, detect, purify…)


Module 2

  • End alginate research
  • Other effects of transcription factor other than colanic acid production
  • How do we detect if we are making our Biobrick?
  • Visualisation is ok – not quantitative enough though
  • Experiment

can we show E.coli coated in calcium phosphate?


Module 3

  • Killing depends on output of module 2
  • Promoter leakiness is a PROBLEM
  • Think about which RE's to work with

there is a delicate balance between methyltransferases and RE's worried about over expressing methytransferase above what is required RE may get there first and degrade DNA

  • Go for an E.coli system and go for a whole RE set

Geoff has EcoRV with RE and methylase on two different plasmids

  • Failsafe

develop a robust system have one methyltransferase that protects against two different RE’s

  • Cutting the gene out from either side after promoter is good enough as it is not functional


Linking Modules

  • Need some sort of timer/delay mechanism (Biobrick potential)
  • External inducers?
  • Need to link by experimentation


General

  • Order essentials ASAP (PKU, strain...)
  • Need to match specifications to details in each module…
  • Gantt chart may help to split areas
  • Plan for wet lab (perhaps 2 people in there by the end of this week?)
  • Think about experiments we can carry out per module
  • Think about modelling (interfaces and some of the modules)
  • Remember – think modular!
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